腹膜透析液对腹膜间皮细胞TLR2和TLR4表达的影响

摘要

目的：观察不同浓度葡萄糖（dextrose）腹膜透析液、艾考糊精（icodextrin）腹膜透析液对人腹膜间皮细胞 Toll 样受体2（TLR2）和 TLR4表达的影响。方法体外培养人腹膜间皮细胞株第5～10代（HMrSV5，DMEM ／F12培养基含10％胎牛血清）用于实验研究。 M TT 检测细胞活力，实验分为5组：阴性对照组、1．50％ dextrose 组、2．50％ dextrose 组、4．25％ dextrose 组、7．5％ icodextrin 组，应用蛋白免疫印迹法（Western blot）检测 TLR2和 TLR4的表达水平。结果不同浓度 dextrose 腹膜透析液作用24 h 对腹膜间皮细胞 TLR2和 TLR4的表达无明显影响。作用48 h ，1．50％ dextrose 组、2．50％ dextrose 组、4．25％ dex-trose 组 TLR2分别降低（5．5±2．8）％、（31．4±7．5）％、（54．9±1．9）％，TLR4分别降低（32．9±17．6）％、（47．7±13．5）％、（66．4±13．5）％；作用72 h ，1．50％ dextrose 组、2．50％ dextrose 组、4．25％ dextrose 组 TLR2表达分别降低（29．4±14．7）％、（38．9±9．9）％、（63．5±16．5）％，TLR4表达分别降低（59．5±16．8）％、（63．1±9．5）％、（79．2±14．0）％；7．5％ icodextrin 组TLR2和 TLR4表达与阴性对照组比较差异无统计学意义（P＞0．05）。结论 dextrose 透析液下调 HM rSV5 TLR2和 TLR4的表达；icodextrin 透析液对 TLR2和 TLR4的表达无显著影响。%Objective To investigate the effect of glucose-based peritoneal dialysis fluids and icodextrin-based peritoneal dial-ysis fluids on the expression of TLR2 and TLR4 on huamn peritoneal mesothelial cells .Methods Human peritoneal mesothelial cell line 5 - 10 generations(HMrSV5) was cultured in DMEM /F12 medium supplemented with 10% (v/v) fetal calf serum (FCS) .Cell viability and cell proliferation were assessed using M TT method .The experiment were divided into 5 different groups :group A (control group) ,1 .5% dextrose group ,2 .5% dextrose group ,4 .25% dextrose group and 7 .5% Lcodextrin group .Icodextrin group (aikau dextrin) ,TLR2 and TLR4 expression were detected by Western blot .Results Treatment with different concentrations of glucose-based peritoneal dialysis fluids for 24 h did not affect the expression of TLR2 and TLR4 protein .In addition ,after stimula-tion for 48 h ,1 .5% dextrose ,2 .5% dextrose ,4 .25% dextrose decreased TLR2 expression by (5 .5 ± 2 .8)% ,(31 .4 ± 7 .5)% , (54 .9 ± 1 .9)% respectively ,TLR4 expression by (32 .9 ± 17 .6)% ,(47 .7 ± 13 .5)% ,(66 .4 ± 13 .5)% respectively .Stimulation for 72 h ,decreased TLR2 expression by (29 .4 ± 14 .7)% ,(38 .9 ± 9 .9)% ,(63 .5 ± 16 .5)% respectively ,TLR4 expression by(59 .5 ± 16 .8)% ,(63 .1 ± 9 .5)% ,(79 .2 ± 14 .0)% respectively .There was no significant change in TLR2 and TLR4 protein expression on 7 .5% icodextrin group .Conclusion Glucose-based peritoneal dialysis fluids ,but not icodextrin-based peritoneal dialysis fluids downregulates expression of TLR2 and TLR4 by HM rSV5 .